Phenyl substituted imidaopyridines and phenyl substituted benzimidazoles

ABSTRACT

Compounds described by the formula (I) or formula (II): (I), (II), or pharmaceutically acceptable salts thereof, are inhibitors of p38 useful in the treatment of inflammatory diseases such as arthritis. Compounds may be selective adenosine A1 antagonists useful in the treatment of neurological disorders such as dementia and depression.

BACKGROUND OF THE INVENTION

Mitogen-activated protein (“MAP”) kinases mediate the surface-to-nucleussignal transduction in a cell. Protein kinases that activate andphosphorylate MAP are known as mitogen-activated protein kinase kinases(“MKK”). One such MKK specifically phosphorylates and activates the p38MAP kinase (“p38”) and is called MKK3. U.S. Pat. Nos. 5,736,381 and5,804,427 describe human mitogen-activated kinase kinase isoforms.International Publication No. 98/00539 describes a human gene encodingan M3-Interacting Protein.

Xia et al., Science, 270:1326-1331(1995) describes the p38 signaltransduction pathway as being activated by proinflammatory cytokines andenvironmental stress. MKK3 is described as being involved in transducingstress signals such as nerve growth factor mediated apoptosis in PC12cells. It is believed that inhibition of p38 activity can provide relieffrom acute and chronic inflammation by blocking production of cytokinessuch as IL-1 and TNF, thereby inhibiting the production ofproinflammatory cytokines such as IL-6 and IL-8. In particular, it isbelieved that p38 inhibitors block the synthesis of TNFα and IL-1β,cytokines, thereby providing relief from inflammatory diseases such asrheumatoid arthritis. Accordingly, it would be desirable to providenovel compounds that are selective and potent inhibitors of the actionof p38.

International Publication No. 97/22704 describes the mitogen-activatedprotein kinase kinase MEK6, which can stimulate phosphorylation andactivation of p38 substrates. International Publication Nos. 95/31451,99/00357 and 98/27098 describe various inhibitors of p38. Nonetheless,there remains a great need to develop inhibitors of the action of p38for various pharmaceutical and therapeutic applications.

The following reviews describe the biochemistry of adenosine receptormodulation and the application to neuropharmacology: Guieu, et al., Gen.Pharmac. 31:553-561(1998), Poulsen and Quinn, Bioorg. Med. Chem.6:619-641(1998) and Williams, Nucleosides Nucleotides10:1087-1099(1991). Adenosine G-protein coupled receptors are located atthe synapses of neurons and on dendrites, and the adenosine A₁ subtypeis primarily distributed in brain tissue. Endogenous adenosine is knownto inhibit the release of many neurotransmitters, excitatory amino acidsand hormones. This phenomenon occurs through the GPCR-mediated blockadeof calcium ion channel effectors, decreasing the influx of calcium intocells of the central or peripheral nervous system. Antagonism of thissedative effect serves to increase the levels of neurotransmitters suchas acetylcholine, dopamine, serotonin, GABA and glutamate, several ofwhich have been successfully targeted in the treatment of neurologicaldisorders by their upregulation. In particular, for instance, adenosineAl antagonism is believed to enhance cognition by the upregulation ofacetylcholine and glutamate, and therefore may have therapeuticapplication to dementias such as Alzheimer's disease. Accordingly, itwould be desirable to provide novel compounds that are selective andpotent antagonists of the action of adenosine with application toneuroscience pharmacology.

International Publication Nos. 01/39777 and 01/40230 describe variousadenosine antagonists with minimal A₁ subtype selectivity. Nonetheless,there remains a great need to develop selective adenosine antagonistsfor various pharmaceutical and therapeutic applications.

SUMMARY OF THE INVENTION

The present invention relates to compounds of the following Formula (I)or (II):

or a pharmaceutically acceptable salt and/or hydrate thereof,

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compounds represented by Formula(I) or Formula (II):

or a pharmaceutically acceptable salt or hydrate thereof, wherein

-   -   the dotted line indicates an optional bond;    -   R¹ is hydrogen, C₁₋₆alkyl- group, C₃₋₆ cycloalkyl- group, aryl        group, or arylC₁₋₆alkyl- group, any of the groups optionally        substituted with 1-6 substituents, each substituent        independently being —OH, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-C(O)—C₀₋₄alkyl-, or halogen;    -   R² is hydrogen, —C(O)—N₃, —NCO, C₁₋₆alkyl- group,        —C(O)(C₀₋₄alkyl) group, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl)        group, —(C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group,        —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —O—C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—(C₀₋₄alkyl)        group, —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl) group, or        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—C₀₋₄alkyl)aryl group, any of the        groups optionally substituted with 1-6 substituents, each        substituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;    -   R³¹, R³², R³³, R³⁴, R³⁵ each independently is hydrogen, halogen,        or C₁₋₆alkyl- group optionally substituted with 1-6        substituents, each substituent independently being —OH,        —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-,        or halogen;    -   n is 0, 1, or 2; and    -   any alkyl is optionally substituted with 1-6 independent        halogen.

In one aspect, the present invention is directed to compoundsrepresented by Formula (I):

or a pharmaceutically acceptable salt or hydrate thereof, wherein

-   -   the dotted line indicates an optional bond;    -   R¹ is hydrogen, C₁₋₆alkyl- group, C₃₋₆ cycloalkyl- group, aryl        group, or arylC₁₋₆alkyl- group, any of the groups optionally        substituted with 1-6 substituents, each substituent        independently being —OH, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-C(O—C₀₋₄alkyl-, or halogen;    -   R² is hydrogen, —C(O)—N₃, —NCO, C₁₋₆alkyl- group,        —C(O)(C₀₋₄alkyl) group, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl)        group, —C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group,        —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —O—C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—C₀₋₄alkyl) group,        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—C₀₋₄alkyl) group, or        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—C₀₋₄alkyl)aryl group, any of the        groups optionally substituted with 1-6 substituents, each        substituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;

R³¹, R³², R³³, R³⁴, R³⁵ each independently is hydrogen, halogen, orC₁₋₆alkyl- group optionally substituted with 1-6 substituents, eachsubstituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;

-   -   n is 0, 1, or2; and    -   any alkyl is optionally substituted with 1-6 independent        halogen.

In an embodiment of this one aspect, the present invention is directedto compounds represented by:

or a pharmaceutically acceptable salt or hydrate thereof, wherein

-   -   R¹ is hydrogen, C₁₋₆alkyl- group, C₃₋₆ cycloalkyl- group, aryl        group, or arylC₁₋₆alkyl- group, any of the groups optionally        substituted with 1-6 substituents, each substituent        independently being —OH, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-C(O)—C₀₋₄alkyl-, or halogen;    -   R² is hydrogen, —C(O)—N₃, —NCO, C₁₋₆alkyl- group,        —C(O)(C₀₋₄alkyl) group, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl)        group, —C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group,        —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —O—C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—(C₀₋₄alkyl)        group, —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl) group, or        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl)aryl group, any of the        groups optionally substituted with 1-6 substituents, each        substituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;    -   R³¹, R³², R³³, R³⁴, R³⁵ each independently is hydrogen, halogen,        or C₁₋₆alkyl- group optionally substituted with 1-6        substituents, each substituent independently being —OH,        —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-,        or halogen;    -   n is 0, 1, or2; and    -   any alkyl is optionally substituted with 1-6 independent        halogen.

In another embodiment of this one aspect, the present invention isdirected to compounds represented by:

or a pharmaceutically acceptable salt or hydrate thereof, wherein

-   -   R¹ is hydrogen, C₁₋₆alkyl- group, C₃₋₆ cycloalkyl- group, aryl        group, or arylC₁₋₆alkyl- group, any of the groups optionally        substituted with 1-6 substituents, each substituent        independently being —OH, —(C₀₋₄alkyl-N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-C(O)—C₀₋₄alkyl-, or halogen;    -   R² is hydrogen, —C(O)-N₃, —NCO, C₁₋₆alkyl- group,        —C(O)(C₀₄alkyl) group, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl)        group, —(C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group,        —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —O—C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—C₀₋₄alkyl) group,        -C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl) group, or        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl)aryl group, any of the        groups optionally substituted with 1-6 substituents, each        substituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋ ₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;    -   R³¹, R³², R³³, R³⁴, R³⁵ each independently is hydrogen, halogen,        or C₁₋₆alkyl- group optionally substituted with 1-6        substituents, each substituent independently being —OH,        —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-,        or halogen;    -   n is 0, 1,or2;and    -   any alkyl is optionally substituted with 1-6 independent        halogen.

In a second aspect, the present invention is directed to compoundsrepresented by formula (II):

or a pharmaceutically acceptable salt or hydrate thereof, wherein

-   -   R¹ is hydrogen, C₁₋₆alkyl- group, C₃₋₆ cycloalkyl- group, aryl        group, or arylC₁₋₆alkyl- group, any of the groups optionally        substituted with 1-6 substituents, each substituent        independently being —OH, —(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-C(O)C₀₋₄alkyl-, or halogen;    -   R² is hydrogen, —C(O)N₃, —NCO, C₁₋₆alkyl- group,        —C(O)(C₀₋₄alkyl) group, —C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl)        group, —(C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group,        —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —C(O)-N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,        —O—C(O)-N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—(C₀₋₄alkyl)        group, —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl) group, or        —C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl)aryl group, any of the        groups optionally substituted with 1-6 substituents, each        substituent independently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl),        C₁₋₄alkyl, C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen;    -   R³¹, R³², R³³, R³⁴, R³⁵ each independently is hydrogen, halogen,        or C₁₋₆alkyl- group optionally substituted with 1-6        substituents, each substituent independently being —OH,        —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-,        or halogen;    -   n is 0, 1, or 2; and    -   any alkyl is optionally substituted with 1-6 independent        halogen.

As used herein, “alkyl” as well as other groups having the prefix “alk”such as, for example, alkoxy, alkanoyl, alkenyl, alkynyl and the like,means carbon chains which may be linear or branched or combinationsthereof. Examples of alkyl groups include methyl, ethyl, propyl,isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl and thelike. “Alkenyl”, “alkynyl” and other like terms include carbon chainscontaining at least one unsaturated C—C bond.

The term “cycloalkyl” means carbocycles containing no heteroatoms, andincludes mono-, bi- and tricyclic saturated carbocycles, as well asfused ring systems. Such fused ring systems can include one ring that ispartially or fully unsaturated such as a benzene ring to form fused ringsystems such as benzofused carbocycles. Cycloalkyl includes such fusedring systems as spirofused ring systems. Examples of cycloalkyl includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, decahydronaphthalene,adamantane, indanyl, indenyl, fluorenyl, 1,2,3,4-tetrahydronaphalene andthe like. Similarly, “cycloalkenyl” means carbocycles containing noheteroatoms and at least one non-aromatic C—C double bond, and includemono-, bi- and tricyclic partially saturated carbocycles, as well asbenzofused cycloalkenes. Examples of cycloalkenyl include cyclohexenyl,indenyl, and the like.

The term “aryl” means an aromatic substituent which is a single ring ormultiple rings fused together. When formed of multiple rings, at leastone of the constituent rings is aromatic. The preferred arylsubstituents are phenyl and naphthyl groups.

The term “cycloalkyloxy” unless specifically stated otherwise includes acycloalkyl group connected by a short C₁₋₂alkyl length to the oxyconnecting atom.

The term “C₀₋₆alkyl” includes alkyls containing 6, 5, 4, 3, 2, 1, or nocarbon atoms. An alkyl with no carbon atoms is a hydrogen atomsubstituent when the alkyl is a terminal group and is a direct bond whenthe alkyl is a bridging group.

The term “hetero” unless specifically stated otherwise includes one ormore O, S, or N atoms. For example, heterocycloalkyl and heteroarylinclude ring systems that contain one or more O, S, or N atoms in thering, including mixtures of such atoms. The hetero atoms replace ringcarbon atoms. Thus, for example, a heterocycloC₅alkyl is a five-memberring containing from 4 to no carbon atoms. Examples of heteroarylsinclude pyridinyl, quinolinyl, isoquinolinyl, pyridazinyl, pyrimidinyl,pyrazinyl, quinoxalinyl, furyl, benzofuryl, dibenzofuryl, thienyl,benzthienyl, pyrrolyl, indolyl, pyrazolyl, indazolyl, oxazolyl,benzoxazolyl, isoxazolyl, thiazolyl, benzothiazolyl, isothiazolyl,imidazolyl, benzimidazolyl, oxadiazolyl, thiadiazolyl, triazolyl, andtetrazolyl. Examples of heterocycloalkyls include azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl,imidazolinyl, pyrolidin-2-one, piperidin-2-one, and thiomorpholinyl.

The term “heteroC₀₋₄alkyl” means a heteroalkyl containing 3, 2, 1, or nocarbon atoms. However, at least one heteroatom must be present. Thus, asan example, a heteroC₀₋₄alkyl having no carbon atoms but one N atomwould be a —NH—if a bridging group and a —NH₂ if a terminal group.Analogous bridging or terminal groups are clear for an O or Sheteroatom.

The term “amine” unless specifically stated otherwise includes primary,secondary and tertiary amines substituted with C₀₋₆alkyl.

The term “carbonyl” unless specifically stated otherwise includes aC₀₋₆alkyl substituent group when the carbonyl is terminal. That is,“carbonyl” means —C(O)—C₀₋₆alkyl unless otherwise stated.

The term “halogen” includes fluorine, chlorine, bromine and iodineatoms.

The term “optionally substituted” is intended to include bothsubstituted and unsubstituted. Thus, for example, optionally substitutedaryl could represent a pentafluorophenyl or a phenyl ring. When a grouphas an optional substituent, that optional substituent can be on any ofthe sites readily determined and understood by chemists. That is, forexample, a substituent on a cyclopropylC₁₋₄alkyl group can be on thecyclopropyl or on the C₁₋₄alkyl. Further, optionally substitutedmultiple moieties such as, for example, alkylaryl are intended to meanthat the aryl and the alkyl groups are optionally substituted. If onlyone of the multiple moieties is optionally substituted then it will bespecifically recited such as “an alkylaryl, the aryl optionallysubstituted with halogen or hydroxyl.”

Compounds described herein contain one or more double bonds and may thusgive rise to cis/trans isomers as well as other conformational isomers.The present invention includes all such possible isomers as well asmixtures of such isomers unless specifically stated otherwise.

Compounds described herein can contain one or more asymmetric centersand may thus give rise to diastereomers and optical isomers. The presentinvention includes all such possible diastereomers as well as theirracemic mixtures, their substantially pure resolved enantiomers, allpossible geometric isomers, and pharmaceutically acceptable saltsthereof. The above Formula I and II are shown without a definitivestereochemistry at certain positions. The present invention includes allstereoisomers of Formula I and II, and pharmaceutically acceptable saltsthereof. Further, mixtures of stereoisomers as well as isolated specificstereoisomers are also included. During the course of the syntheticprocedures used to prepare such compounds, or in using racemization orepimerization procedures known to those skilled in the art, the productsof such procedures can be a mixture of stereoisomers.

Unless specifically stated otherwise or indicated by a bond symbol (dashor double dash), the connecting point to a recited group will be on theright-most stated group. That is, for example, a phenylalkyl group isconnected to the main structure through the alkyl and the phenyl is asubstituent on the alkyl.

The compounds of the present invention are useful in variouspharmaceutically acceptable salt forms. The term “pharmaceuticallyacceptable salt” refers to those salt forms which would be apparent tothe pharmaceutical chemist. i.e., those which are substantiallynon-toxic and which provide the desired pharmacokinetic properties,palatability, absorption, distribution, metabolism or excretion. Otherfactors, more practical in nature, which are also important in theselection, are cost of the raw materials, ease of crystallization,yield, stability, hygroscopicity and flowability of the resulting bulkdrug. Conveniently, pharmaceutical compositions may be prepared from theactive ingredients in combination with pharmaceutically acceptablecarriers.

The pharmaceutically acceptable salts of the compounds of Formula I andII include conventional non-toxic salts or quarternary ammonium salts ofthe compounds of Formula I and II formed e.g. from non-toxic inorganicor organic acids. For example, non-toxic salts include those derivedfrom inorganic acids such as hydrochloric, hydrobromic, sulfuric,sulfamic, phosphoric, nitric and the like; and the salts prepared fromorganic acids such as acetic, propionic, succinic, glycolic, stearic,lactic, malic, tartaric, citric, ascorbic, pamoic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic, trifluoroacetic and the like.

The pharmaceutically acceptable salts of the present invention can besynthesized by conventional chemical methods. Generally, the salts areprepared by reacting the free base or acid with stoichiometric amountsor with an excess of the desired salt-forming inorganic or organic acidor base, in a suitable solvent or solvent combination.

The compounds of the present invention may have asymmetric centers andoccur as racemates, racemic mixtures, and as individual diastereomers.All such isomers, including optical isomers, being included in thepresent invention.

The invention described herein also includes a pharmaceuticalcomposition which is comprised of a compound described by Formula (I) or(II), or a pharmaceutically acceptable salt thereof, in combination witha pharmaceutically acceptable carrier. The pharmaceutical compositionsof the present invention comprise a compound represented by Formula I orII (or pharmaceutically acceptable salts thereof) as an activeingredient, a pharmaceutically acceptable carrier and optionally othertherapeutic ingredients or adjuvants. Such additional therapeuticingredients include, for example, i) Leukotriene receptor antagonists,ii) Leukotriene biosynthesis inhibitors, iii) corticosteroids, iv) H1receptor antagonists, v) beta 2 adrenoceptor agonists, vi) COX-2selective inhibitors, vii) statins, viii) non-steroidalanti-inflammatory drugs (“NSAID”), and ix) M2/M3 antagonists.

The invention described herein also includes a method of treatingarthritis which is comprised of administering to a mammalian patient inneed of such treatment a compound described by Formula (I) or (II), or apharmaceutically acceptable salt thereof, in an amount which iseffective to treat arthritis. The invention includes methods of treatingarthritis by administering to a mammalian patient in need of suchtreatment a compound described by Formula (I) or (II), or apharmaceutically acceptable salt thereof, in combination or incoadministration with a COX-2 inhibitor.

The invention described herein also includes a method of treating acytokine mediated disease in a mammal, comprising administering to amammalian patient in need of such treatment an amount of a compounddescribed by Formula (I) or (II), or a pharmaceutically acceptable saltthereof, in an amount which is effective to treat said cytokine mediateddisease.

Of particular interest is a method of treating inflammation in amammalian patient in need of such treatment, which is comprised ofadministering to said patient an anti-inflammatory effective amount of acompound described by Formula (I) or (II), or a pharmaceuticallyacceptable salt thereof.

Another method which is of particular interest is a method of treating acytokine mediated disease as described herein wherein the disease isosteoporosis.

Another method which is of particular interest is a method of treating acytokine mediated disease as described herein wherein the disease isnon-osteoporotic bone resorption.

Yet another method which is of particular interest is a method oftreating a cytokine mediated disease as described herein wherein thedisease is Crohn's disease.

This invention also relates to a method of treating arthritis in amammal in need such treatment, which comprises administering to saidmammal an amount of a compound of Formula I or II which is effective fortreating arthritis. Such method includes the treatment of rheumatoid andosteoarthritis.

When administered to a patient for the treatment of athritis, the dosageused can be varied depending upon the type of arthritis, the age andgeneral condition of the patient, the particular compound administered,the presence or level of toxicity or adverse effects experienced withthe drug, and other factors. A representative example of a suitabledosage range is from as low as about 0.01 mg/kg to as high as about 100mg/kg. However, the dosage administered is generally left to thediscretion of the physician.

This invention also relates to a method of inhibiting the action of p38in a mammal in need thereof, which comprises administering to saidmammal an effective amount of a compound described by Formula (I) or(II), or a pharmaceutically acceptable salt thereof, to inhibit saidaction of p38, down to normal levels, or in some cases to subnormallevels, so as to ameliorate, prevent or treat the disease state.

The compounds of Formula I or II can be used in the prophylactic ortherapeutic treatment of disease states in mammals which are exacerbatedor caused by excessive or unregulated cytokines, more specifically IL-1,IL-6, IL-8 or TNF.

The compounds of this invention demonstrates efficacy in the assaysdescribed below. Efficacy is shown in the assays by results of less than10 μM. Advantageously, compounds have results less than 1μM. Even moreadvantageously, compounds have results less than 0.01 μM. Still moreadvantageously, compounds have results in the assays of less than 0.01μM. Because the compounds of Formula I or II inhibit cytokines, such asIL-1, IL-6, IL-8 and TNF, by inhibiting the action of p38 the compoundsare useful for treating diseases in which cytokine presence or activityis implicated, such as rheumatoid arthritis, rheumatoid spondylitis,osteoarthritis, gouty arthritis and other arthritic conditions.

The compounds described by Formula (I) or (II), or a pharmaceuticallyacceptable salt thereof, are also useful to treat other disease statesmediated by excessive or unregulated TNF production or activity. Suchdiseases include, but are not limited to sepsis, septic shock, endotoxicshock, gram negative sepsis, toxic shock syndrome, adult respiratorydistress syndrome, cerebral malaria, chronic pulmonary inflammatorydisease, silicosis, pulmonary sarcoidosis, bone resorption diseases,such as osteoporosis, reperfusion injury, graft v. host rejection,allograft rejection, fever, myalgia due to infection, cachexia secondaryto infection or malignancy, cachexia secondary to acquired immunedeficiency syndrome (AIDS), AIDS, ARC (AIDs related complex), keloidformation, scar tissue formation, Crohn's disease, ulcerative colitis,pyresis, AIDS and other viral infections, such as cytomegalovirus (CMV),influenza virus, and the herpes family of viruses such as Herpes Zosteror Simplex I and II.

The compounds described by Formula (I) or (II), or a pharmaceuticallyacceptable salt thereof, are also useful topically in the treatment ofinflammation such as in the treatment of rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions; inflamed joints, eczema, psoriasis or otherinflammatory skin conditions such as sunburn; inflammatory eyeconditions including conjunctivitis; pyresis, pain and other conditionsassociated with inflammation.

The compounds described by Formula (I) or (II), or a pharmaceuticallyacceptable salt thereof, are also useful in treating diseasescharacterized by excessive IL-8 activity. These disease states includepsoriasis, inflammatory bowel disease, asthma, cardiac and renalreperfusion injury, adult respiratory distress syndrome, thrombosis andglomerulonephritis.

The invention thus includes a method of treating psoriasis, inflammatorybowel disease, asthma, cardiac and renal reperfusion injury, adultrespiratory distress syndrome, thrombosis and glomerulonephritis, in amammal in need of such treatment, which comprises administering to saidmammal a compound described by Formula (I) or (II), or apharmaceutically acceptable salt thereof, in an amount which iseffective for treating said disease or condition.

When administered to a patient for the treatment of a disease in which acytokine or cytokines are implicated, the dosage used can be varieddepending upon the type of disease, the age and general condition of thepatient, the particular compound administered, the presence or level oftoxicity or adverse effects experienced with the drug, and otherfactors. A representative example of a suitable dosage range is from aslow as about 0.01 mg/kg to as high as about 100 mg/kg. However, thedosage administered is generally left to the discretion of thephysician.

The methods of treatment are preferably carried out by delivering thecompound of Formula I or II parenterally. The term ‘parenteral’ as usedherein includes intravenous, intramuscular, or intraperitonealadministration. The subcutaneous and intramuscular forms of parenteraladministration are generally preferred. The instant invention can alsobe carried out by delivering the compound of Formula I or IIsubcutaneously, intranasally, intrarectally, transdermally orintravaginally.

The compounds of Formula I or II may also be administered by inhalation.By ‘inhalation’ is meant intranasal and oral inhalation administration.Appropriate dosage forms for such administration, such as an aerosolformulation or a metered dose inhaler, may be prepared by conventiontechniques.

The invention also relates to a pharmaceutical composition comprising acompound of Formula I or II and a pharmaceutically acceptable carrier.The compounds of Formula I or II may also be included in pharmaceuticalcompositions in combination with a second therapeutically activecompound.

The pharmaceutical carrier employed may be, for example, either a solid,liquid or gas. Exemples of solid carriers include lactose, terra alba,sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate,stearic acid and the like. Exemples of liquid carriers are syrup, peanutoil, olive oil, water and the like. Examples of gaseous carriers includecarbon dioxide and nitrogen.

Similarly, the carrier or diluent may include time delay material wellknown in the art, such as glyceryl monostearate or glyceryl distearate,alone or with a wax.

A wide variety of pharmaceutical dosage forms can be employed. If asolid dosage is used for oral administration, the preparation can be inthe form of a tablet, hard gelatin capsule, troche or lozenge. Theamount of solid carrier will vary widely, but generally will be fromabout 0.025 mg to about 1 g. When a liquid dosage form is desired fororal administration, the preparation is typically in the form of asyrup, emulsion, soft gelatin capsule, suspension or solution. When aparenteral dosage form is to be employed, the drug may be in solid orliquid form, and may be formulated for administration directly or may besuitable for reconstitution.

Topical dosage forms are also included. Examples of topical dosage formsare solids, liquids and semi-solids. Solids would include dustingpowders, poultices and the like. Liquids include solutions, suspensionsand emulsions. Semi-solids include creams, ointments, gels and the like.

The amount of a compound of Formula I or II used topically will, ofcourse, vary with the compound chosen, the nature and severity of thecondition, and can be varied in accordance with the discretion of thephysician. A representative, topical, dose of a compound of Formula I orII is from as low as about 0.01 mg to as high as about 2.0 g,administered one to four, preferably one to two times daily.

The active ingredient may comprise, for topical administration, fromabout 0.001% to about 10% w/w.

Drops according to the present invention may comprise sterile ornon-sterile aqueous or oil solutions or suspensions, and may be preparedby dissolving the active ingredient in a suitable aqueous solution,optionally including a bactericidal and/or fungicidal agent and/or anyother suitable preservative, and optionally including a surface activeagent. The resulting solution may then be clarified by filtration,transferred to a suitable container which is then sealed and sterilizedby autoclaving or maintaining at 98-100° C. for half an hour.Alternatively, the solution may be sterilized by filtration andtransferred to the container aseptically. Examples of bactericidal andfungicidal agents suitable for inclusion in the drops are phenylmercuricnitrate or acetate (0.002%), benzalkonium chloride (0.01%) andchlorhexidine acetate (0.01%). Suitable solvents for the preparation ofan oily solution include glycerol, diluted alcohol and propylene glycol.

Lotions according to the present invention include those suitable forapplication to the skin or eye. An eye lotion may comprise a sterileaqueous solution optionally containing a bactericide and may be preparedby methods similar to those for the preparation of drops. Lotions orliniments for application to the skin may also include an agent tohasten drying and to cool the skin, such as an alcohol or acetone,and/or a moisturizer such as glycerol or an oil such as castor oil orarachis oil.

Creams, ointments or pastes according to the present invention aresemi-solid formulations of the active ingredient for externalapplication. They may be made by mixing the active ingredient infinely-divided or powdered form, alone or in solution or suspension inan aqueous or non-aqueous liquid, with a greasy or non-greasy base. Thebase may comprise hydrocarbons such as hard, soft or liquid paraffin,glycerol, beeswax, a metallic soap; a mucilage; an oil of natural originsuch as almond, corn, arachis, castor or olive oil; wool fat or itsderivatives, or a fatty acid such as stearic or oleic acid together withan alcohol such as propylene glycol or macrogels. The formulation mayincorporate any suitable surface active agent such as an anionic,cationic or non-ionic surfactant such as sorbitan esters orpolyoxyethylene derivatives thereof. Suspending agents such as naturalgums, cellulose derivatives or inorganic materials such as silicas, andother ingredients such as lanolin may also be included.

Assays

Protein Expression and Purification.

Murine p38 containing the FLAG epitope tag was expressed in DrosophilaS2 cells under transcriptional control of a copper-induciblemetallothionein promoter. Expression of recombinant p38 was induced bytreating transfected cells with 1 mM CuSO₄ for 4 hours. To generateactive recombinant murine p38, CuSO₄-treated S2 cells were stimulated 10minutes prior to harvest with 400 mM NaCl, 2 mM Na₃ VO₄, and 100 μg/Lokadaic acid. Cell pellets were washed with phosphate-buffered saline, 2mM Na₃ VO₄, and lysed in 20 mM Tris HCl, pH 7.5, 120 mM NaCl, 1% TritonX-100, 2mM EDTA, 20 mM NaF, 4 mM Na₃ VO₄, 2 mM Prefabloc SC (BoehringerMannheim). Cell lysates were centrifuged for 10 min at 13,00 ×g, andactivated, recombinant murine p38 was immunoaffinity purified from thelysate by column chromatography through anti-FLAG M2 resin (Kodak) thathad been equilibrated with lysis buffer. After loading the extract theresin was washed with 10 column volumes of lysis buffer, 10 columnvolumes buffer A (10 mM Tris HCl, pH 7.5, 500 mM NaCl, 20% glycerol) and10 column volumes of buffer B (10 mM Tris HCl pH 7.5, 150 mM NaCl, 20%glycerol). The fusion protein was eluted in buffer B containing 100μg/mL FLAG peptide (Kodak).

The N-terminal 115 amino acids of ATF-2 was expressed in E. coli as afusion protein with glutathione-S-transferase. The fusion protein waspurified over glutathione agarose according to standard procedures(Pharmacia).

p38 Kinase Assay.

p38 kinase assays were performed in a reaction volume of 100 μL in a96-well plate, at 30° for 45-1200 min under the following conditions: 25mM Hepes, pH 7.4, 10 mMmgCl₂, 2 mM β-glycerolphosphate, 2 mM DTT, 5 μMATP, 10 μCi [γ-³³P]-ATP and ˜2 μM GST-ATF2. Serial dilutions ofcompounds were added to each reaction in 2 μL DMSO. 2 μL of DMSO wasadded to the last row of each reaction plate as the no inhibitor controlfor each inhibitor titration. The reaction was terminated with an equalvolume of a stop solution containing 100 mM EDTA and 15 mM sodiumpyrophosphate. PVDF filter plates (MAIPNOB50, Millipore) were pre-wetwith methanol and washed with the stop solution. 50 μL aliquots from asingle reaction were applied to the filter under vacuum, and the filterwas washed twice with 75 mM phosphoric acid. The filter plates werecounted in a scintillation counter (Top Count, Packard) and the percentinhibition at each compound concentration is determined.

TNF-α Release Assay.

Blood was obtained from healthy volunteers by venipuncture using sodiumheparin as an anti-coagulant. Peripheral blood mononuclear cells (PBMCs)were isolated using Lymphocyte Separation Medium (ICN) according tomanufacturers specifications. Isolated PBMCs were washed 3 times withHBSS and diluted to a density of 2×10⁶ cells/mL in RPMI+5% autologoushuman serum. 50 μL of the serial dilutions of inhibitor were added towells of a 96-well tissue culture plate followed by addition of 100 μLof PBMCs and then 50 μL of RPMI complete medium containing 400 ng/mLLPS. A control well of cells without compound but with LPS (maximalstimulation control) and one without compound and without LPS(background control) were included in each titration. The cells wereincubated for 16 hours in a humidified incubator at 37° C., 5% CO₂.Supernatants were then harvested and TNF-α levels were quantified byimmunoassay using commercial reagents (R&D, Inc).

Human Adenosine A₁ Receptor Binding Assay

Human brain cortex membrane preparations were purchased from ABS, Inc(Wilmington, Del.) and were treated with 2 U/mL adenosine deaminase for15 min on ice, prior to use. The assay was conducted in MilliporeMultiscreen MAFC filter plates (Millipore Corp., MA), using 50 mMTris/HCl, pH 7.4 as binding buffer. The adenosine A₁ selectiveantagonist, (“DPCPX“) 3H-cyclopentyl-1,3 -dipropylxanthine,8-[dipropyl-2,3-3H(N)] (NEN, Boston, Mass.) was used as the radioligandat a final concentration of 0.6 nM. Dilutions of compounds were preparedin DMSO at 100× the desired assay concentration. Typically, finalcompound concentration ranged from 10 μM -500 pM. Unlabeled8-cyclopentyl-1,3-dipropylxanthine (Sigma, Saint Louis, Mo.) was titeredas a positive control. 100 μg of human cortex membranes was added toeach well of the assay and the reaction was allowed to incubate for 1 hat rt. Wells in which inhibitors were omitted served as 0% inhibition.Wells in which 1 μM 8-cyclopentyl-1,3-dipropylxanthine was presentserved as 100% inhibition. At the end of the incubation period, theplates were filtered and washed twice with 100 μL of ice cold bindingbuffer. After transfer to adapter plates (Packard, Downers Grove, Ill.),50 μL Ready Safe scintillation cocktail (Beckman, Fullerton, Calif.) wasadded. Plates were sealed and placed on a shaker for 1 min, and countedon a Topcount (Packard). Percent inhibition was calculated for each welland IC₅₀ values were determined based on a four parameter fit algorithm.

Alternate Human Adenosine A₁ Receptor Binding Assay

Alternatively, adenosine A₁ radioligand binding was performed asfollows. Human recombinant CHO cells were used with 1 nM ³H DPCPX asligand. The vehicle used was 1% DMSO, the incubation buffer used was 20mM HEPES, pH 7.4, 10 mM MgCl₂, 100 mM NaCl, and the incubationconditions were 90 min at 25° C. A non-specific ligand (referencecompound) 100 μM R(−)-PIA (N6-(R-phenylisopropyl)adenosine) was used,and specific binding was 85%, B=2.7 pmol/mg protein, K_(d)=1.4 nM.

Human Adenosine A_(2A) Receptor Binding Assay

Adenosine A_(2A) radioligand binding was performed as follows. Humanrecombinant HEK-293 cells were used with 0.05 μM ³H2-[[p-(2-carboxyethyl)phenethyl]amino]-5′-N-ethylcarboxamidoadenosine(“CGS-21680”) as ligand. The vehicle used was 1% DMSO, the incubationbuffer used was 50 mM Tris-HCl, pH 7.4, 10 mM MgCl₂, 1 mM EDTA, 2U/mLadenosine deaminase, and the incubation conditions were 90 min at 25° C.The non-specific ligand (reference compound) used was 50 μM5′-N-ethylcarboxamidoadenosine (“NECA”), and specific binding was 85%,B_(max)=7 pmol/mg protein, K_(d)=0.064 μM.

Human Adenosine A_(2B) Receptor Binding Assay

Adenosine A_(2B) radioligand binding was performed as follows. Humanrecombinant HEK-293 cells were used with 9 nM ³H DPCPX as ligand. Thevehicle used was 1% DMSO, the incubation buffer used was 10 mM HEPES, pH7.4, 1 mM EDTA, 0.1 mM benzamidine, 2U/mL adenosine deaminase, and theincubation conditions were 80 min at 25° C. The non-specific ligand(reference compound) used was 10 μM DPCPX, and specific binding was 60%,B_(max)=0.96 pmol/mg protein, K_(d)=0.04 μM.

Rat Adenosine A₃ Receptor Binding Assay

Adenosine A₃ radioligand binding was performed as follows. Ratrecombinant EBNA cells were used with 1 nM ¹²⁵I AB-MECA as ligand. Thevehicle used was 1% DMSO, the incubation buffer used was 50 mM Tris-HCl,pH 7.4, 1 mM EDTA, 10 mM MgCl₂, 1.5 U/mL adenosine deaminase added freshat the time of assay, and the incubation conditions were 4 h at 25° C.The non-specific ligand (reference compound) used was 100 μM R(−)-PIA,and specific binding was 90%, B_(max)=1.3 pmol/mg protein, K_(d)=1.3 nM.

Rat Adenosine A₁ Tissue Assay.

An adenosine A₁ tissue assay was performed as follows to determineantagonist versus agonist functional activity. Wistar rat (ca. 275 g)vas deferens were used with the adenosine A₁ agonist reference compound,N6-(cyclohexyl)adenosine (“CHA”) (0.3 μM, 100%) and the adenosine A₁antagonist reference compound, DPCPX (10 nM, 87%). The vehicle used was0.1% DMSO, the incubation buffer used was Krebs at pH 7.4, and theincubation time was 5 min at 32° C. The administration volume was 10 μL,the bath volume was 10 mL, and the time of assessment was 5 min using anisometric (gram changes) quantitation method. The criteria forfunctional agonism was a ≧50% reduction of neurogenic twitch relative toCHA response. The criteria for functional antagonism was a ≧50%inhibition of CHA-induced relaxation.

The compounds of this invention demonstrated efficacy in the aboveassays by results of less than 10 μM. Advantageous compounds had resultsless than 1 μM. Even more advantageous compounds had results less than0.1 μM. Still more advantageous compounds had results in the assays ofless than 0.01 μM. In the above assays, compounds of this inventiondemonstrated significant functional adenosine A₁ antagonism (ca. 60%) inan IC₅₀ range of 1 nM to 100 nM. In the above assays, compounds of thisinvention demonstrated 500-fold binding selectivity for the adenosine A₁receptor over the adenosine A_(2A) receptor subtype. In the aboveassays, compounds of this invention demonstrated >1000-fold bindingselectivity for the adenosine A₁ receptor over the A_(2B) and A₃adenosine receptor subtypes. Certain compounds of this inventiondemonstrated 20-fold selectivity for p38 kinase inhibition overadenosine A₁ receptor binding in the above assays. Certain compounds ofthis invention demonstrated a range of 10-fold to 200-fold selectivityfor adenosine A₁ receptor binding over p38 kinase inhibition in theabove assays.

Thus, the compounds of this invention are effective inhibitors ofcytokines—particularly p38 and TNF-alpha. Accordingly, the compounds ofthis invention are effective to treat inflammation in a mammalianpatient in need of such treatment by administering to the patient ananti-inflammatory effective amount of a compound of this invention.

As a result, the compounds of this invention are also effective fortreating rheumatoid arthritis, osteoarthritis, endotoxemia, toxic shocksyndrome, inflammatory bowel disease, tuberculosis, atherosclerosis,muscle degeneration, cachexia, psoriatic arthritis, Reiter's syndrome,rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis oracute synovitis by administering an effective amount of a compound ofthis invention.

Further, as a result, the compounds of this invention are also effectivefor treating rheumatoid arthritis, rheumatoid spondylitis,osteoarthritis, gouty arthritis, sepsis, septic shock, endotoxic shock,gram negative sepsis, toxic shock syndrome, adult respiratory distresssyndrome, cerebral malaria, chronic pulmonary inflammatory disease,silicosis, pulmonary sarcosis, bone resorption diseases, reperfusioninjury, graft v. host rejection, allograft rejection, fever, myalgia dueto infection, cachexia secondary to infection or malignancy, cachexiasecondary to acquired immune deficiency syndrome (AIDS), AIDS relatedcomplex (ARC), keloid formation, scar tissue formation, Crohn's disease,ulcerative colitis or pyresis by adminstering an effective amount of acompound of this invention.

Further, as a result, the compounds of this invention are effectuve fortreating osteoporosis in a mammalian patient in need of such treatment.

Further, as a result, the compounds of this invention are effective fortreating bone resorption in a mammalian patient in need of suchtreatment.

Further, as a result, the compounds of this invention are effective fortreating Crohn's disease in a mammalian patient in need of suchtreatment.

Also as a result, the compounds of this invention are effective fortreating neurodegenerative disease, Parkinson's disease, anxiety,psychosis, schizophrenia, and substance abuse.

Further, as a result, the compounds of this invention are effective fortreating pain and migraine.

Further, as a result, the compounds of this invention are also effectivefor treating stroke and cerebrovascular disease.

Further, as a result, the compounds of this invention are effective asantidementia, antidepressant, antianxiety, antipsychotic, anticatalepsy,antiparkinsonian, anxiolytic, nootropic, analgesic, or psychostimulentcompounds. The compounds are also effective as a therapeutic forcerebral circulation.

Further, as a result, the compounds of this invention can be used forcognitive enhancement, for their antidepressant action, their cerebralvasodilating action, and for their action of increasing cerebral bloodflow.

Furthermore, the selective adenosine A₁ antagonism properties of thecompounds of this invention leads to the compounds being effective inthe treatment and prevention of depression and dementia (eg. Alzheimer'sdisease, cerebrovascular dementia, and dementia accompanying Parkinson'sdisease).

The compounds of the invention are prepared by the following reactionschemes. All substituents are as defined above unless indicatedotherwise.

The following examples illustrate the preparation of some of thecompounds of the invention and are not to be construed as limiting theinvention disclosed herein.

COMPOUND Ia was prepared from the commercially available methyl4-fluorobenzoyl acetate. To a solution of methyl 4-fluorobenzoyl acetate(10 g, 51.0 mmol) in CH₂Cl₂ (130 mL) at 0° C. was added solidtetrabutylammonium tribromide (26 g, 53.6 mmol). The reaction mixturewas maintained at 0° C. for 2 h and then slowly warmed to 23° C. andmaintained for 1 h. The orange reaction mixture was then partitionedbetween NaHCO_(3(aq)) and CH₂Cl₂, the organic phase washed thrice withNaHCO_(3(aq)), then dried over anhydrous sodium sulfate and concentratedin vacuo. The crude product was vacuum pumped for 30 min, diluted intoanhydrous ethanol (250 mL) and treated with commercially available2-aminopyridine (24 g, 255 mmol). The reaction mixture was warmed to 60°C. and maintained for 14 h, cooled to rt, partitioned betweenNaHCO_(3(aq)) and CHCl₃, the organic phase dried over anhydrous sodiumsulfate and concentrated in vacuo. The crude material was purified byflash chromatography (Biotage 65M, SiO₂, hexane to 20% acetone-hexanegradient elution) to provide COMPOUND Ia which was characterized by ¹HNMR, HPLC and mass spectrometry (m/z: 271 (M⁺+1)).

COMPOUND Ia (9.5 g, 35.2 mmol) was diluted into dry THF (176 mL) andN,O-dimethylhydroxylamine hydrochloride (10.3 g, 105.6 mmol) was added.The mixture was cooled to −10° C. and a 2 molar solution ofisopropylmagnesium chloride in THF (106 mL, 211.1 mmol) was added undernitrogen. The reaction mixture was maintained at −10° C. for 1 h andthen quenched into water. The mixture was then partitioned betweenNaHCO_(3(aq)) and CH₂Cl₂, the organic phase dried over anhydrous sodiumsulfate and concentrated in vacuo. The crude material was purified byflash chromatography (Biotage 65M, SiO₂, 50% ethyl acetate-hexane toethyl acetate gradient elution) to provide the Weinreb amide productwhich was characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 300(M⁺+1)).

This material (7.75 g, 25.9 mmol) was diluted into dry THF (150 mL),cooled to 0° C. and treated with a 3 molar solution of methylmagnesiumbromide in diethyl ether (26 mL, 77.8 mmol) under nitrogen. The reactionmixture was maintained at 0° C. for 30 min, quenched into water,partitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudematerial can either be purified by flash chromatography (Biotage, SiO₂,hexane to 30% acetone-hexane gradient elution) or used in the next stepwithout purification. The highly pure methyl ketone COMPOUND IIa wascharacterized by ¹H NMR, HPLC and mass spectrometry (m/z: 255 (M⁺+1)).

COMPOUND IIa (6.4 g, 25.2 mmol) was diluted into dry THF (250 mL),cooled to −78° C., and treated with a 1 molar solution of lithiumbis(trimethylsilyl)amide in THF (38 mL, 37.8 mmol) under nitrogen. Themixture was maintained at −78° C. for 30 min and then treated slowlywith t-butyl bromoacetate (19 mL, 125.9 mmol) at −78° C. under nitrogen.The reaction mixture was maintained at −78° C. for 1 h and then slowlywarmed to 23° C. over 1 h. The mixture was then quenched into water,partitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in-vacuo. The crudet-butyl ketoester product was then diluted into dry CH₂Cl₂ (180 mL),cooled to 0° C. and treated with trifluoroacetic acid (63 mL). Thereaction mixture was maintained at 0° C. for 2 h, warmed to 23° C. for 2h and then concentrated in vacuo. The crude residue was then dilutedinto dry methanol, cooled to 0° C., and treated with excess hydrogenchloride gas for 3-5 min. The reaction mixture was maintained at 0° C.for 1 h, and partitioned between NaHCO_(3(aq)) and CHCl₁₃. The organicphase was dried over anhydrous sodium sulfate and concentrated in vacuo.The crude material was purified by flash chromatography (Biotage 65M,SiO₂, hexane to 30% acetone-hexane gradient elution) to provide COMPOUNDIIIa which was characterized by ¹H NMR, HPLC and mass spectrometry (m/z:327 (M⁺+1)).

EXAMPLE 1

COMPOUND IIIa (32 mg, 0.098 mmol) was combined with sodium acetate (241mg, 2.94 mmol) and commercially available cyclohexylhydrazinehydrochloride (296 mg, 1.96 mmol). Glacial acetic acid (3.5 mL) andwater (1.5 mL) were added, and the reaction mixture was refluxed at 130°C. for 20 h. The mixture was cooled to rt, partitioned between aqueous2N NaOH and CH₂Cl₂, ensuring an aqueous pH >9, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudematerial was purified by preparative centrifugal thin layerchromatography (Chromatotron, 4 mm SiO₂, 20% to 70% ethyl acetate-hexanegradient elution) to provide semi-pure EXAMPLE 1 which was furtherpurified by preparative reverse phase HPLC (Gilson, YMC C₁₈, 90% H₂O(0.05% TFA)-10% CH₃CN (0.05% TFA) 1 min isocratic then 9 min gradientelution to 100% CH₃CN (0.05% TFA). The product eluent was reduced involume to aqueous, treated with solid NaHCO_(3(aq)) ensuring an aqueouspH >9, partitioned into CH₂Cl₂ and concentrated in vacuo to provideEXAMPLE 1 which was characterized by ¹H NMR, HPLC and mass spectrometry(m/z: 391 (M⁺+1)).

EXAMPLES 2-21

The following compounds were prepared under conditions similar to thosedescribed above, culminating in the synthesis of EXAMPLE 1. Thedifferent groups represented below as R¹ were introduced by thesubstitution of the appropriate commercially available hydrazine orhydrazine salt in place of cyclohexylhydrazine hydrochloride as shownabove in Scheme 1. The different phenyl groups represented below as Ar²were introduced by the substitution of the appropriate β-ketoester(benzoyl acetates were commercially available or prepared by literaturemethods known to those skilled in the art) in place of methyl4-fluorobenzoyl acetate as shown in Scheme 1. The following exampleswere characterized by HPLC and mass spectrometry, and in most cases,additionally by ¹H NMR and/or high resolution mass spectrometry.

MS (m/z) EX. R^(1 Group) Ar^(2 Group) (M⁺ + 1) 2 2-Hydroxyethyl4-Fluorophenyl 353 3 2,2,2-Trifluoroethyl 4-Fluorophenyl 391 4 H4-Fluorophenyl 309 5 Benzyl 4-Fluorophenyl 399 6 Isopropyl4-Fluorophenyl 351 7 2-Chlorophenyl 4-Fluorophenyl 419 8 3-Chlorophenyl4-Fluorophenyl 419 9 2-Chlorophenyl 2-Chlorophenyl 436 10 Cyclohexyl2,4-Difluorophenyl 409 11 2-Chlorophenyl 3-(Trifluoromethyl)phenyl 46912 Cyclohexyl 3-(Trifluoromethyl)phenyl 441 13 2-Chlorophenyl2-Chloro-4--fluorophenyl 454 14 2,6-Dichlorophenyl2-Chloro-4-fluorophenyl 488 15 2-Tolyl 2-Chloro-4-fluorophenyl 433 162,6-Dichlorophenyl 2,3-Dichlorophenyl 505 17 2-Chlorophenyl2,3-Dichlorophenyl 470 18 2-Tolyl 2,3-Dichlorophenyl 450 192-(Trifluoromethyl) 2,3-Dichlorophenyl 504 phenyl 20 2-Tolyl2,4-Difluorophenyl 417 21 2,6-Dichlorophenyl 2,4-Difluorophenyl 472

EXAMPLE 22

EXAMPLE 1 (2 mg, 0.005 mmol) was combined with copper (II) chloride (34mg, 0.256 mmol) and diluted into dry acetonitrile (0.2 mL). The reactionmixture was refluxed at 85° C. for 74 h. The mixture was cooled to rt,concentrated in vacuo, partitioned between water and CH₂Cl₂, treatedwith concentrated ammonium hydroxide, the organic phase dried overanhydrous sodium sulfate and concentrated in vacuo. The crude productwas purified by preparative thin layer chromatography (500 micron SiO₂,20×10 cm, 60% ethyl acetate-hexane) to provide EXAMPLE 22 which wascharacterized by ¹H NMR, HPLC and mass spectrometry (m/z: 389 (M⁺+1)).

EXAMPLES 23-37

The following pyridazinones were prepared from their respectivedihydropyridazinones under oxidative conditions similar to thosedescribed for the synthesis of EXAMPLE 22 as shown in Scheme 1. Thefollowing examples were characterized by HPLC and mass spectrometry, andin most cases, additionally by ¹H NMR and/or high resolution massspectrometry.

MS (m/z) EX. R¹ Group Ar² Group (M⁺ + 1) 23 2,2,2-Trifluoroethyl4-Fluorophenyl 389 24 2,6-Dichlorophenyl 4-Fluorophenyl 452 252-Chlorophenyl 2-Chlorophenyl 434 26 Cyclohexyl 2,4-Difluorophenyl 40727 2-Chlorophenyl 3-(Trifluoromethyl)phenyl 467 28 Cyclohexyl3-(Trifluoromethyl)phenyl 439 29 2,6-Dichlorophenyl3-(Trifluoromethyl)phenyl 502 30 2-Chlorophenyl 2-Chloro-4-fluorophenyl452 31 2,6-Dichlorophenyl 2-Chloro-4-fluorophenyl 486 32 2-Tolyl2-Chloro-4-fluorophenyl 431 33 2,6-Dichlorophenyl 2,3-Dichlorophenyl 50334 2-Tolyl 2,3-Dichlorophenyl 448 35 2-(Trifluoromethyl)2,3-Dichlorophenyl 502 phenyl 36 2-Tolyl 2,4-Difluorophenyl 415 372,6-Dichlorophenyl 2,4-Difluorophenyl 470

COMPOUND IVa was prepared in an analogous manner to the t-butylketoester precursor of COMPOUND IIIa except that2-amino4-(hydroxymethyl)pyridine (commercially available from CBResearch, New Castle, Del.) and 2-chloro4-fluorobenzoyl acetate (seeEXAMPLES 2-21) were used in place of 2-aminopyridine and 4-fluorobenzoylacetate respectively. The product COMPOUND IVa was purified on SiO₂using 20% ethyl acetate-hexane eluent and characterized by ¹H NMR, HPLCand mass spectrometry (m/z: 547 (M⁺+1)).

COMPOUND IVa (690 mg, 1.26 mmol) was combined with pyridiniump-toluenesulfonate (1.3 g, 5.18 mmol) and diluted into MeOH (13 mL). Thereaction mixture was maintained at 23° C. for 1 h and then concentratedin vacuo. The crude residue was purified by preparative centrifugal thinlayer chromatography (Chromatotron, 4 mm SiO₂, hexane to 30%acetone-hexane to 1:9:90 NH₄OH-MeOH—CHCl₃ 3 step gradient elution) toprovide 500 mg (92%) of alcohol which was characterized by ¹H NMR, HPLCand mass spectrometry (m/z: 433 (M⁺+1)). This material (200 mg, 0.462mmol) was diluted into dry THF (8mL), treated with1,8-diazabicyclo[5.4.0]undec-7-ene (0.065 mL, 0.647 mmol) followed bydiphenylphosphoryl azide (0.119 mL, 0.554 mmol), and the reactionmixture was maintained at 23° C. for 14 h. The reaction mixture waspartitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The cruderesidue was purified by preparative centrifugal thin layerchromatography (Chromatotron, 2 mm SiO₂, 20% ethyl acetate-hexaneisocratic elution) to provide 140 mg (66%) of azide which wascharacterized by ¹H NMR. This material (140 mg, 0.306 mmol) was dilutedinto THF (8 mL), treated with water (0.220 mL, 12.2 mmol) followed bytriphenylphosphine (240 mg, 0.918 mmol), and the reaction mixture wasmaintained at 23 ° C. for 14 h. The reaction mixture was partitionedbetween NaHCO_(3(aq)) and 30% isopropanol-chloroform, the organic phasedried over anhydrous sodium sulfate and concentrated in vacuo. The cruderesidue was purified by preparative centrifugal thin layerchromatography (Chromatotron, 2 mm SiO₂, chloroform to 20%methanol-chloroform to 50% methanol-chloroform to 1:9:90NH₄OH-MeOH—CHCl₃ 4 step gradient elution) to provide COMPOUND Va whichwas characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 432(M⁺+1)).

COMPOUND Va (29 mg, 0.067 mmol) was diluted into CH₂Cl₂ (1.3 mL),treated with diisopropylethylamine (0.035 mL, 0.198 mmol), cooled to 0°C., and methanesulfonyl chloride (0.008 mL, 0.099 mmol) added undernitrogen. The reaction mixture was maintained at 0° C. for 3 h,partitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The cruderesidue was purified by preparative reverse phase HPLC (Gilson, YMC C₁₈,90% H₂O (0.05% TFA) - 10% CH₃CN (0.05% TFA) 1 min isocratic then 9 mingradient elution to 100% CH₃CN (0.05% TFA). The product eluent wasreduced in volume to aqueous, treated with solid NaHCO_(3(aq)) ensuringan aqueous pH >9, partitioned into CH₂Cl₂ and concentrated in vacuo toprovide the sulfonamide t-butyl ketoester which was characterized by ¹HNMR, HPLC and mass spectrometry (m/z: 510 (M⁺+1)). This material (7 mg,0.0138 mmol) was then diluted into dry CH₂Cl₂ (0.140 mL), cooled to 0°C. and treated with trifluoroacetic acid (0.140 mL). The reactionmixture was maintained at 0° C. for 2 h, warmed to 23° C. for 2 h andthen concentrated in vacuo. The crude residue was then diluted into drymethanol, cooled to 0° C., and treated with excess hydrogen chloride gasfor 1 min. The reaction mixture was maintained at 0° C. for 1 h,partitioned between NaHCO_(3(aq)) and CHCl₃, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudeproduct sulfonamide methyl ketoester COMPOUND VIa was used directly inthe next cyclization reaction.

EXAMPLE 38

Crude COMPOUND VIa (6 mg, 0.0138 mmol) was combined with sodium acetate(36 mg, 0.435 mmol) and 2-tolylhydrazine hydrochloride (46 mg, 0.29mmol). Glacial acetic acid (1 mL) and water (0.2 mL) were added, and thereaction mixture was refluxed at 150° C. for 16 h. The mixture wascooled to rt, partitioned between aqueous 2N NaOH and CH₂Cl₂ ensuring anaqueous pH >9, the organic phase dried over anhydrous sodium sulfate andconcentrated in vacuo. The crude material was purified by preparativethin layer chromatography (SiO₂, 250 micron, 20×20 cm, 30% THF-hexanethen 1:3:96 NH₄OH-MeOH—CHCl₃ then acetonitrile, 3 step gradient elution)to provide EXAMPLE 38 which was characterized by ¹H NMR, HPLC and massspectrometry (m/z: 540 (M⁺+1)).

Commercially available 2-chloro4-aminopyridine (5 g, 38.8 mmol) wascombined with trityl chloride (14 g, 50.4 mmol), catalyticdimethylaminopyridine (DMAP, 470 mg, 3.88 mmol), diluted into drymethylene chloride (130 mL) and treated with triethylamine (17 mL, 116.3mmol). The reaction mixture was maintained at 23° C. for 15 h,partitioned between NaHCO_(3(aq)) and CHCl₃, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudematerial was purified by plug flash chromatography using a sinteredglass funnel with vacuum (SiO₂, 13×13 cm, hexane to 30% ethylacetate-hexane gradient elution). The 4-trityl-protected amine wascharacterized by ¹H NMR and HPLC. This material (6 g, 0.016 mol) wascombined with cesium carbonate (26.4 g, 0.081 mol),2-(di-t-butylphosphino)biphenyl (1.9 g, 0.0065 mol), Pd₂(dba)₃ (3 g,0.0032mol), diluted into dry DME (100 mL) and treated with benzophenoneimine (27 mL, 0.162 mol). The reaction mixture was refluxed at 100° C.for 15 h under nitrogen, cooled to rt, partitioned between NaHCO_(3(aq))and CHCl₃, the organic phase dried over anhydrous sodium sulfate andconcentrated in vacuo. The crude intermediate imine adduct was thencombined with sodium acetate (27 g, 0.324 mol) and methoxylaminehydrochloride (20 g, 0.243 mol), diluted into dry methanol (160 mL) andmaintained at 23° C. for 1 h. The reaction mixture was partitionedbetween NaHCO_(3(aq)) and CH₂Cl₂, the organic phase dried over anhydroussodium sulfate and concentrated in vacuo. The crude material waspurified by flash chromatography (Biotage 65M, Sio₂, 50% ethylacetate-hexane then 1:9:90 NH₄OH-MeOH—CHCl₃ 2 step gradient elution) toprovide the 4-trityl-protected COMPOUND VIIa which was characterized by¹H NMR and HPLC. This intermediate (1.5 g, 4.3 mmol) was deprotected bydilution into methylene chloride (14 mL) and treatment withtrifluoroacetic acid (4.3 mL) at 23° C. for 2 h. The reaction mixturewas then quenched into water, solid sodium chloride and solid sodiumbicarbonate were added, the mixture partitioned into 30%isopropanol-chloroform, and the organic phase was dried over anhydroussodium sulfate and concentrated in vacuo. The crude material waspurified by flash chromatography (Biotage 65M, SiO₂, 100% hexane to 100%chloroform gradient elution followed by 1:9:90 and then 3:27:90NH₄OH-MeOH—CHCl₃ gradient elution) to provide COMPOUND VIIa which wascharacterized by ¹H NMR and HPLC.

COMPOUND VIIIa was prepared in an analogous manner to COMPOUND Ia exceptthat 1.5 molar equivalents of COMPOUND VIIa was used in place of 5 molarequivalents of 2-aminopyridine, and anhydrous dioxane was substitutedfor ethanol as the reaction solvent. The reaction mixture was warmed to60° C. and maintained for 14 h, cooled to rt, partitioned betweenNaHCO_(3(aq)) and 30% isopropanol-CHCl₃, the aqueous exhaustivelyextracted, and the combined organic phases dried over anhydrous sodiumsulfate and concentrated in vacuo. The crude product was purified bypreparative thin layer chromatography (500 micron SiO₂, 20×20 cm, 30%acetone-hexane) to provide COMPOUND VIIIa which was characterized by ¹HNMR. It was anticipated that COMPOUND VIIIa could be transformed intoamino-substituted EXAMPLES related to and generated from the chemistryshown in Scheme 1 upon minor synthetic modifications known to thoseskilled in the art.

COMPOUND IXa was prepared from commercially available maleic anhydride(9.1 g, 0.093 mol) and ortho-tolylhydrazine hydrochloride (10 g, 0.063mol) which were combined and diluted into water (154 mL), followed bythe addition of concentrated HCl (20 mL). The reaction mixture wasrefluxed at 110° C. for 15 h, cooled to rt, filtered and the precipitatewashed with toluene. The solid product COMPOUND IXa was dried in vacuoand characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 203(M⁺+1)).

COMPOUND IXa (3 g, 14.85 mmol) was combined with P(O)Br₃ (10 g, 34.84mmol) in a sealed tube and the reaction mixture heated at 130° C. for 3h. The mixture was cooled to rt, poured into ice, partitioned betweenwater and ethyl acetate, and the organic phase dried over anhydroussodium sulfate and concentrated in vacuo. The crude material waspurified by flash chromatography (Biotage 65M, SiO₂, hexane to 60% ethylacetate-hexane gradient elution) to provide the dark orange solidproduct COMPOUND Xa XIV which was characterized by HPLC and massspectrometry (m/z: 265 (M⁺+1)).

Commercially available 3-nitro-4-aminobenzoic acid (10 g, 54.9 mmol) wasdiluted into dry THF (133 mL), treated with LiOH—H₂O (670 mg, 27.9 mmol)at 23° C. for 30 min, and dimethyl sulfate (670 mg, 5.3 mmol) was thenadded. The reaction mixture was heated at 75° C. for 13 h undernitrogen. The mixture was cooled to rt, partitioned betweenNaHCO_(3(aq)) and ethyl acetate, and the organic phase dried overanhydrous sodium sulfate and concentrated in vacuo. The crude materialwas purified by flash chromatography (Biotage 65M, SiO₂, hexane to 20%acetone-hexane gradient elution) to provide COMPOUND XIa which wascharacterized by ¹H NMR, HPLC and mass spectrometry (m/z: 197 (M⁺+1)).

COMPOUND XIa (1 g, 5.1 mmol) was combined with COMPOUND Xa (1 g, 3.77mmol), cesium carbonate (290 mg, 0.89 mmol), Pd₂(dba)₃ (430 mg, 0.47mmol), Xanthphos (570 mg, 0.99 mmol) and diluted into dry degassed DME(40 mL). The reaction mixture was refluxed at 90° C. for 20 h undernitrogen in a sealed tube, cooled to rt, filtered and concentrated invacuo. The crude material was purified by flash chromatography (Biotage40M, SiO₂, hexane to 20% ethyl acetate-hexane to ethyl acetate gradientelution) to provide the dark yellow solid product COMPOUND XIIa whichwas characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 381(M⁺+1)).

COMPOUND XIIa (600 mg, 1.58 mmol) was diluted into dry methanol (87 mL)and methylene chloride (10 mL), heated to homogeneity, and then thesolution was cooled to −30° C. To this cooled reaction mixture was addeda methanol solution of catalytic Raney nickel (washed once with waterand twice with methanol), a hydrogen atmosphere was introduced viaballoon, purged thrice, and the reaction mixture was stirred vigerouslyat −30° C. for 3 h excluding light. The reaction mixture was filteredthrough a pad of Celite, quickly washed with methylene chloride andconcentrated in vacuo. The crude material was purified by flashchromatography (Biotage 40M, SiO₂, hexane to CH₂Cl₂ to 5% MeOH—CH₂Cl₂gradient elution) to provide COMPOUND XIIIa which was characterized by¹H NMR, HPLC and mass spectrometry (m/z: 351 (M⁺+1)).

EXAMPLE 39

COMPOUND XIIIa (290 mg, 0.83 mmol) was diluted into dry nitrobenzene (21mL), treated with excess molecular sieves, and2-chloro4-fluorobenzaldehyde (145 mg, 0.91 mmol) was added. The reactionmixture was heated in a sealed tube at 170° C. for 15 h, and thenitrobenzene was then distilled away at 110° C. The crude product waspurified by flash chromatography (Biotage 40M, SiO₂, hexane to CH₂Cl₂ to5% MeOH—CH₂Cl₂ gradient elution) to provide EXAMPLE 39 which wascharacterized by HPLC and mass spectrometry (m/z: 489 (M⁺+1)).

EXAMPLE 40

EXAMPLE 39 (250 mg, 0.51 mmol) was diluted into (3:1:1) THF-MeOH—H₂O (5mL), treated with a 1N solution of aqueous LiOH (2 mL), and the reactionmixture was maintained at 23° C. for 4 h. The reaction mixture wasneutralized with a 1N solution of aqueous HCl (2 mL), and chloroform (15mL) was added. The solution was dried with excess anhydrous sodiumsulfate and concentrated in vacuo to provide EXAMPLE 40 which wascharacterized by HPLC and mass spectrometry (m/z: 475 (M⁺+1)).

EXAMPLE 41

EXAMPLE 40 (50 mg, 0.105 mmol) was diluted into dry CH₂Cl₂ (0.5 mL),treated with triethylamine (0.046 mL, 0.316 mmol), diphenylphosphorylazide (0.033 mL, 0.158 mmol), and the reaction mixture was maintained at23° C. for 14 h. The reaction mixture was then purified directly bypreparative thin layer chromatography (SiO₂, 1000 micron, 20×20 cm, 50%ethyl acetate-hexane) to provide EXAMPLE 41 which was characterized byHPLC and mass spectrometry (m/z: 500 (M⁺+1)).

EXAMPLE 42

EXAMPLE 41 (20 mg, 0.040 mmol) was diluted into dry toluene (0.8 mL) andthe reaction mixture was heated at 80° C. for 4 h. The intermediateisocyanate was characterized by HPLC and mass spectrometry (m/z: 472(M⁺+1)) directly from the reaction mixture and was not isolated. Theisocyanate EXAMPLE 42 was used directly in the next reaction to formcarbamate EXAMPLE 43.

EXAMPLE 43

EXAMPLE 42 (5 mg, 0.0106 mmol) in toluene (0.2 mL) was treated with dryMeOH (0.002 mL, 0.053 mmol) and maintained at 23° C. for 14 h. Thereaction mixture was then partitioned between NaHCO_(3(aq)) and 30%isopropanol-chloroform, and the organic phase dried over anhydroussodium sulfate and concentrated in vacuo to provide EXAMPLE 43 which wascharacterized by ¹H NMR, HPLC and mass spectrometry (m/z: 504 (M⁺+1)).

EXAMPLE 44

EXAMPLE 44 was prepared as in EXAMPLE 43 by the addition ofdimethylamine to isocyanate EXAMPLE 42 in toluene. The purified productwas characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 517(M⁺+1)).

COMPOUND XIa (1 g, 5.10 mmol) was diluted into dry THF (70 mL), cooledto 0° C., and treated with lithium aluminum hydride (250 mg, 6.57 mmol)after which the reaction mixture was maintained at 0° C. for 15 min. Themixture was partitioned between water and ethyl acetate, and the organicphase dried over anhydrous sodium sulfate and concentrated in vacuo. Thecrude material was then diluted into DMF (30 mL), treated with imidazole(1.6 g, 24 mmol), tert-butyldiphenylsilyl chloride (1.8 g, 12 mmol) andmaintained at 23° C. for 15 h. The reaction mixture was partitionedbetween water and diethyl ether, and the organic phase dried overanhydrous sodium sulfate and concentrated in vacuo. The crude materialwas purified by flash chromatography (SiO₂, 15% acetone-hexane) toprovide COMPOUND XIVa which was characterized by ¹H NMR and HPLC.

EXAMPLE 45

COMPOUND XIVa was manipulated through the same transformations describedfor the synthesis of EXAMPLE 39 to provide the silyl-protected EXAMPLE45. The tert-butyldiphenylsilyl ether (400 mg, 0.569 mmol) was dilutedinto dry THF (15 mL), cooled to 0° C., and treated with a 1M THFsolution of tetrabutylammonium fluoride (0.63 mL, 0.626 mmol) afterwhich the reaction mixture was maintained at 0° C. for 2 h. The mixturewas partitioned between water and ethyl acetate, and the organic phasedried over anhydrous sodium sulfate and concentrated in vacuo. The crudematerial was purified by flash chromatography (SiO₂, acetone-hexane) toprovide EXAMPLE 45 which was characterized by HPLC and mass spectrometry(m/z: 465 (M⁺+1)).

EXAMPLE 46

EXAMPLE 45 (25 mg, 0.054 mmol) was diluted into dry CH₂Cl₂ (1 mL) andtreated with Dess-Martin reagent (34 mg, 0.081 mmol) at 23° C. underargon. The reaction mixture was maintained at 23° C. for 2 h,partitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudeproduct was purified by preparative thin layer chromatography (SiO₂, 2plates, 1000 micron, 20×20 cm, 5% MeOH—CHCl₃) to provide EXAMPLE 46which was characterized by ¹H NMR, HPLC and mass spectrometry (m/z: 463(M⁺+1)).

EXAMPLE 47

EXAMPLE 46 (19 mg, 0.041 mmol) was diluted into dry CH₂Cl₂ (1 mL),treated with N,N-diisopropylethylamine (0.023 mL, 0.123 mmol), a 2M THFsolution of dimethylamine (0.031 mL, 0.062 mmol), sodiumtriacetoxyborohydride (17 mg, 0.082 mmol), and the reaction mixture wasmaintained at 23° C. for 15 h. The reaction mixture was then partitionedbetween NaHCO_(3(aq)) and CH₂Cl₂, the organic phase dried over anhydroussodium sulfate and concentrated in vacuo to provide EXAMPLE 47 which wascharacterized by ¹H NMR, HPLC and mass spectrometry (m/z: 492 (M⁺+1)).

EXAMPLE 48

EXAMPLE 45 (50 mg, 0.108 mmol) was diluted into dry CH₂Cl₂ (0.6 mL) andTHF (1.1 mL), treated with a 1M CH₂Cl₂ solution of PBr₃ (0.3 mL, 0.323mmol), and the reaction mixture was maintained at 23° C. for 15 h underargon. The reaction mixture was then partitioned between NaHCO_(3(aq))and CH₂Cl₂, the organic phase dried over anhydrous sodium sulfate andconcentrated in vacuo. The crude product was purified by preparativethin layer chromatography (SiO₂, 4 plates, 1000 micron, 20×20 cm, 30%acetone-hexane) to provide EXAMPLE 48 which was characterized by ¹H NMR,HPLC and mass spectrometry (m/z: 527 (M⁺+1)).

EXAMPLE 49

EXAMPLE 48 (38 mg, 0.072 mmol) was diluted into dry CH₂Cl₂ (1.5 mL),treated with NaSMe (15 mg, 0.217 mmol), and the reaction mixture wasmaintained at 23° C. for 15 h under argon. The reaction mixture was thenpartitioned between NaHCO_(3(aq)) and CH₂Cl₂, the organic phase driedover anhydrous sodium sulfate and concentrated in vacuo. The crudeproduct was purified by preparative thin layer chromatography (SiO₂, 2plates, 1000 micron, 20×20 cm, 30% ethyl acetate-hexane) to provideEXAMPLE 49 which was characterized by ¹H NMR, HPLC and mass spectrometry(m/z: 495 (M⁺+1)).

EXAMPLE 50

EXAMPLE 49 (21 mg, 0.043 mmol) was diluted into MeOH (1 mL), and treateddropwise with a water solution (0.4 mL) of oxone (56 mg, 0.089 mmol).The reaction mixture was maintained at 23° C. for 2 h, partitionedbetween water and CH₂Cl₂, the organic phase dried over anhydrous sodiumsulfate and concentrated in vacuo. The crude product was purified bypreparative thin layer chromatography (SiO₂, 1000 micron, 20×20 cm, 40%acetone-hexane) to provide EXAMPLE 50 which was characterized by ¹H NMR,HPLC and mass spectrometry (m/z: 527 (M⁺+1)).

EXAMPLE 51

EXAMPLE 51 was prepared from EXAMPLE 45 following the procedure inCOMPOUND Va for the generation of the amine from the intermediatealcohol. The purified product was characterized by HPLC and massspectrometry (m/z: 464 (M⁺+1)).

EXAMPLE 52

EXAMPLE 52 was prepared from EXAMPLE 51 following the procedure inCOMPOUND VIa for the generation of the methylsulfonamide. The purifiedproduct was characterized by ¹H NMR, HPLC and mass spectrometry (m/z:542 (M⁺+1)).

EXAMPLE 53

EXAMPLE 53 was prepared from EXAMPLE 51 following the procedure inCOMPOUND VIa for the generation of the methylsulfonamide, butsubstituting toluenesulfonyl chloride for methanesulfonyl chloride. Thepurified product was characterized by HPLC and mass spectrometry (m/z:618 (M⁺+1)).

4-Amino-5-nitro-2-methylthiopyrimidine (50 mg, 0.269 mmol), prepared byliterature methods known to those skilled in the art, was diluted intoDME (2 mL), combined with COMPOUND Xa (110 mg, 0.403 mmol) and degassedvia a stream of nitrogen bubbled through the mixture. Cesium carbonate(88 mg, 0.269 mmol), xanthphos (20 mg, 0.035 mmol) and Pd₂(dba)₂ (15 mg,0.016 mmol) were added sequentially, and the reaction mixture was heatedat 95° C. for 15 h under argon. The reaction mixture was filteredthrough a pad of celite, washed with DME and concentrated in vacuo. Thecrude product was purified by preparative thin layer chromatography (4plates, SiO₂, 1000 micron, 20×20 cm, 50% ethyl acetate-hexane) toprovide COMPOUND XVa which was characterized by ¹H NMR, HPLC and massspectrometry (m/z: 371 (M⁺+1)).

COMPOUND XVa (50 mg, 0.135 mmol) was diluted into CH₂Cl₂ (4 mL), treatedwith catalytic palladium on carbon, evacuated and flushed with hydrogengas via a double balloon, and stirred at rt for 14 h under a positivepressure of hydrogen. The reaction mixture was filtered through a pad ofcelite, washed with methylene chloride and concentrated in vacuo. Thecrude product was purified by preparative thin layer chromatography (2plates, SiO₂, 1000 micron, 20×20 cm, 50% ethyl acetate-hexane) toprovide COMPOUND XVIa which was characterized by HPLC and massspectrometry (m/z: 341 (M⁺+1)).

EXAMPLE 54

COMPOUND XVIa (16 mg, 0.047 mmol) was diluted into dry nitrobenzene (15mL), treated with excess molecular sieves, and 2,4-difluorobenzaldehyde(0.007 mL, 0.065 mmol) was added. The reaction mixture was heated in asealed tube at 170° C. for 15 h, and the nitrobenzene was then distilledaway at 110° C. The crude product was purified by preparative thin layerchromatography (2 plates, Sio₂, 1000 micron, 20×20 cm, 10% MeOH—CH₂Cl₂)to provide EXAMPLE 54 which was characterized by HPLC and massspectrometry (m/z: 463 (M⁺+1)).

EXAMPLE 55

EXAMPLE 54 (7 mg, 0.015 mmol) was diluted into methanol (0.3 mL),treated dropwise with a solution of oxone (18 mg, 0.032 mmol) in water(0.15 mL), and the reaction mixture was stirred at 23° C. for 2 h. Thereaction mixture was then partitioned between water and CH₂Cl₂, theorganic phase dried over anhydrous sodium sulfate and concentrated invacuo. The crude product, EXAMPLE 55, was characterized by massspectrometry (m/z: 495 (M⁺+1)).

EXAMPLE 56

EXAMPLE 55 (3.5 mg, 0.007 mmol) was diluted into DMSO (1 mL), bubbledwith ammonia gas for 5 min, and the reaction mixture was heated in apressure tube at 100° C. for 1.5 h. The DMSO was then distilled away at100° C. under a stream of nitrogen. The crude residue was purified bypreparative thin layer chromatography (SiO₂, b 250 micron, 20×20 cm, 50%ethyl acetate-hexane) to provide EXAMPLE 56 which was characterized by¹H NMR, HPLC and mass spectrometry (m/z: 432 (M⁺+1)).

EXAMPLE 57

EXAMPLE 57 was prepared as in EXAMPLE 56 by the addition ofdimethylamine to sulfone EXAMPLE 55 in DMSO. The purified product wascharacterized by HPLC and mass spectrometry (m/z: 460 (M⁺+1)).

1. A compound represented by formula (I) or formula (II):

or a pharmaceutically acceptable salt or hydrate thereof, wherein thedotted line indicates an optional bond; R¹ is hydrogen, C₁₋₆alkyl-group, C₃₋₆ cycloalkyl- group, aryl group, or arylC₁₋₆alkyl- group, anyof the groups optionally substituted with 1-6 substituents, eachsubstituent independently being —OH,—(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₄alkyl, C₁₋₆alkoxy,C₁₋₆alkyl-C(O)—C₀₋₄alkyl-, or halogen; R² is hydrogen, —C(O)—N₃, —NCO,C₁₋₆alkyl- group, —C(O)(C₀₋₄alkyl) group,—(C₀₋₄alkyl)-N(C₀₋₄alkyl)(C₀₋₄alkyl) group,—(C₀₋₄alkyl)-S(O)_(n)—(C₀₋₄alkyl) group, —S(O)₂—N(C₀₋₄alkyl)(C₀₋₄alkyl)group, —C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,—N(C₀₋₄alkyl)-C(O)—N(C₀₋₄alkyl)(C₀₋₄alkyl) group,—O—C(O—)N(C₀₋₄alkyl)(C₀₋₄alkyl) group, —C(O)—O—C₀₋₄alkyl) group,—C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—C₀₋₄alkyl) group, or—C₀₋₆alkyl-N(C₀₋₄alkyl)-S(O)₂—(C₀₋₄alkyl)aryl group, any of the groupsoptionally substituted with 1-6 substituents, each substituentindependently being —OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₄alkyl,C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, or halogen; R³¹, R³², R³³, R³⁴, R³⁵each independently is hydrogen, halogen, or C₁₋₆alkyl- group optionallysubstituted with 1-6 substituents, each substituent independently being—OH, —N(C₀₋₄alkyl)(C₀₋₄alkyl), C₁₋₆alkoxy, C₁₋₆alkyl-CO—C₀₋₄alkyl-, orhalogen; n is 0, 1, or2; and any alkyl is optionally substituted with1-6 independent halogen.
 2. The compound according to claim 1,represented by formula (I) or a pharmaceutically acceptable saltthereof.
 3. The compound according to claim 2,

or a pharmaceutically acceptable salt thereof.
 4. The compound accordingto claim 2, represented by

R¹ Group Ar² Group 2-Hydroxyethyl 4-Fluorophenyl 2,2,2-Trifluoroethyl4-Fluorophenyl H 4-Fluorophenyl Benzyl 4-Fluorophenyl Isopropyl4-Fluorophenyl 2-Chlorophenyl 4-Fluorophenyl 3-Chlorophenyl4-Fluorophenyl 2-Chlorophenyl 2-Chlorophenyl Cyclohexyl2,4-Difluorophenyl 2-Chlorophenyl 3-(Trifluoromethyl)phenyl Cyclohexyl3-(Trifluoromethyl)phenyl 2-Chlorophenyl 2-Chloro-4-fluorophenyl2,6-Dichlorophenyl 2-Chloro-4-fluorophenyl 2-Tolyl2-Chloro-4-fluorophenyl 2,6-Dichlorophenyl 2,3-Dichlorophenyl2-Chlorophenyl 2,3-Dichlorophenyl 2-Tolyl 2,3-Dichlorophenyl2-(Trifluoromethyl) 2,3-Dichlorophenyl phenyl 2-Tolyl 2,4-Difluorophenyl2,6-Dichlorophenyl 2,4-Difluorophenyl

or a pharmaceutically acceptable salt thereof.
 5. The compound accordingto claim 2, represented by

R¹ Group Ar² Group 2,2,2-Trifluoroethyl 4-Fluorophenyl2,6-Dichlorophenyl 4-Fluorophenyl 2-Chlorophenyl 2-ChlorophenylCyclohexyl 2,4-Difluorophenyl 2-Chlorophenyl 3-(Trifluoromethyl)phenylCyclohexyl 3-(Trifluoromethyl)phenyl 2,6-Dichlorophenyl3-(Trifluoromethyl)phenyl 2-Chlorophenyl 2-Chloro-4-fluorophenyl2,6-Dichlorophenyl 2-Chloro-4-fluorophenyl 2-Tolyl2-Chloro-4-fluorophenyl 2,6-Dichlorophenyl 2,3-Dichlorophenyl 2-Tolyl2,3-Dichlorophenyl 2-(Trifluoromethyl) 2,3-Dichlorophenyl phenyl 2-Tolyl2,4-Difluorophenyl 2,6-Dichlorophenyl 2,4-Difluorophenyl

or a pharmaceutically acceptable salt thereof.
 6. The compound accordingto claim 1, represented by formula (II) or a pharmaceutically acceptablesalt thereof.
 7. The compound according to claim 6, represented by

or a pharmaceutically acceptable salt thereof.
 8. A pharmaceuticalcomposition comprising a compound in accordance with claim 1 incombination with a pharmaceutically acceptable carrier.
 9. A method oftreating a inflammation in a mammalian patient in need of suchtreatment, which is comprised of administering to said patient ananti-inflammatory effective amount of a compound as described inclaim
 1. 10. A method of treating rheumatoid arthritis, osteoarthritis,endotoxemia, toxic shock syndrome, inflammatory bowel disease,tuberculosis, atherosclerosis, muscle degeneration, cachexia, psoriaticarthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumaticarthritis, rubella arthritis or acute synovitis by administering aneffective amount of a compound as described in claim
 1. 11. A method oftreating rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis,gouty arthritis, sepsis, septic shock, endotoxic shock, gram negativesepsis, toxic shock syndrome, adult respiratory distress syndrome,cerebral malaria, chronic pulmonary inflammatory disease, silicosis,pulmonary sarcosis, bone resorption diseases, reperfusion injury, graftv. host rejection, allograft rejection, fever, myalgia due to infection,cachexia secondary to infection or malignancy, cachexia secondary toacquired immune deficiency syndrome (AIDS), AIDS related complex (ARC),keloid formation, scar tissue formation, Crohn's disease, ulcerativecolitis or pyresis by adminstering an effective amount of a compound asdescribed in claim
 1. 12. A method of treating osteoporosis in amammalian patient in need of such treatment, which is comprised ofadministering to said patient an effective amount of a compound asdescribed in claim
 1. 13. A method of treating bone resorption in amammalian patient in need of such treatment, which is comprised ofadministering to said patient an effective amount of a compound asdescribed in claim
 1. 14. A method of treating Crohn's disease in amammalian patient in need of such treatment which is comprised ofadministering to said patient an effective amount of a compound asdescribed in claim
 1. 15. A method of treating dementia,neurodegeneraton, or Parkinson's disease in a mammalian patient in needof such treatment which is comprised of administering to said patient aneffective amount of a compound as described in claim
 1. 16. A method oftreating depression, anxiety, psychosis, schizophrenia, or substanceabuse in a mammalian patient in need of such treatment which iscomprised of administering to said patient an effective amount of acompound as described in claim
 1. 17. A method of treating pain ormigraine in a mammalian patient in need of such treatment which iscomprised of administering to said patient an effective amount of acompound as described in claim
 1. 18. A method of treating stroke andcerebrovascular disease in a mammalian patient in need of such treatmentwhich is comprised of administering to said patient an effective amountof a compound as described in claim
 1. 19. A process for making apharmaceutical composition comprising combining a compound of claim 1and a pharmaceutically acceptable carrier.